Genotoxic damage in Solea senegalensis exposed to sediments from the Sado Estuary (Portugal): effects of metallic and organic contaminants

Juvenile Solea senegalensis (Senegalese sole) were exposed to freshly collected sediments from three sites of the Sado Estuary (West-Portuguese coast) in 28-day laboratory assays in order to assess the ecological risk from sediment contaminants, by measuring two genotoxicity biomarkers in peripheral blood: the percentage of Erythrocyte Nuclear Abnormalities (ENA) by use of an adaptation of the micronucleus test, and the percentage of DNA strand-breakage (DNA-SB) with the Comet assay. Sediments were surveyed for metallic (Cr, Ni, Cu, Zn, As, Cd and Pb) and organic (PAHs (polycyclic aromatic hydrocarbons), PCBs (polychlorinated biphenyls) and DDTs (dichloro-diphenyl-trichloroethane)) contaminants. Sediments from site A (farthest from hotspots of contamination) were found to be the least contaminated and weaker inducers of genotoxic damage, whereas sediments from sites B (urban influence) and C (affected by industrial effluents and agricultural runoffs) were responsible for a very significant increase in both ENA and DNA-SB, site B being most contaminated with metals and site C mainly with organic pollutants, especially PAHs and PCBs . Analysis of genotoxic effects showed a strong correlation between the concentrations of PAHs and PCBs and both biomarkers at sampling times T(14) and T(28), while the amounts of Cu, As, Cd and Pb were less strongly correlated, and at T(28) only, with ENA and DNA-SB. These results show that organic contaminants in sediment are stronger and faster acting genotoxic stressors. The results also suggest that metals may have an inhibitory effect on genotoxicity when interacting with organic contaminants, at least during early exposure. ENA and DNA-SB do not show a linear relationship, but a strong correlation exists between the overall increase in genotoxicity caused by exposure to sediment, confirming that they are different, and possibly non-linked effects that respond similarly to exposure. Although the Comet assay showed enhanced sensitivity, the two analyses are complementary and suitable for the biomonitoring of sediment contaminants in a benthic species like S. senegalensis.     

Comparison of two extraction procedures for the assessment of sediment genotoxicity: implication of polar organic compounds

Four sediment samples (Va?ne Airport VA, Va?ne Center VC, Va?ne North VN and Reference North RN) were collected in the Berre lagoon (France). Sediments were analyzed for polycyclic aromatic hydrocarbons (PAHs) by use of pressurized fluid extraction with a mixture of hexane/dichloromethane followed by HPLC with fluorescence detection analysis. Organic pollutants were also extracted with two solvents for subsequent evaluation of their genotoxicity: a hexane/dichloromethane mixture intended to select non-polar compounds such as PAHs, and 2-propanol intended to select polar contaminants. Sediment extracts were assessed by the Salmonella/microsome mutagenicity test with Salmonella typhimurium TA98+S9 mix and YG1041±S9 mix. Extracts were also assessed for their DNA-damaging activity and their clastogenic/aneugenic properties by the comet assay and the micronucleus test with Chinese Hamster ovary (CHO) cells. The PAH concentrations were 611ngg(-1)dw, 1341ngg(-1) dw, 613ngg(-1)dw and 482ngg(-1)dw for VA, VC, VN and RN, respectively. Two genotoxic profiles were observed, depending on the extraction procedure. All the non-polar extracts were mutagenic for TA98+S9 mix, and VA, VC, VN sediment samples exerted a significant DNA-damaging and clastogenic activity in the presence of S9 mix. All the polar extracts appeared mutagenic for TA98+S9 mix and YG104±S9 mix, and VA, VC, VN were genotoxic and clastogenic both with and without S9 mix. These results indicate that the genotoxic and mutagenic activities mainly originated from PAHs in the non-polar extracts, while these activities came from other genotoxic contaminants, such as aromatic amines and nitroarenes, in the polar extracts. This study focused on the important role of uncharacterized polar contaminants such as nitro-PAHs or aromatic amines in the global mutagenicity of sediments. The necessity to use appropriate extraction solvents to accurately evaluate the genotoxic hazard of aquatic sediments is also highlighted.     

In vivo Comet assay on isolated kidney cells to distinguish genotoxic carcinogens from epigenetic carcinogens or cytotoxic compounds

The objective of this study was to determine the ability of the alkaline in vivo Comet assay (pH > 13) to distinguish genotoxic carcinogens from epigenetic carcinogens when performed on freshly isolated kidney cells and to determine the possible interference of cytotoxicity by assessing DNA damage induced by renal genotoxic, epigenetic or toxic compounds after enzymatic isolation of kidney cells from OFA Sprague–Dawley male rats. The ability of the Comet assay to distinguish (1) genotoxicity versus cytotoxicity and (2) genotoxic versus non-genotoxic (epigenetic) carcinogens, was thus investigated by studying five known genotoxic renal carcinogens acting through diverse mechanisms of action, i.e. streptozotocin, aristolochic acids, 2-nitroanisole, potassium bromate and cisplatin, two rodent renal epigenetic carcinogens: d-limonene and ciclosporine and two nephrotoxic compounds: streptomycin and indomethacin. Animals were treated once with the test compound by the appropriate route of administration and genotoxic effects were measured at the two sampling times of 3–6 and 22–26 h after treatment. Regarding the tissue processing, the limited background level of DNA migration observed in the negative control groups throughout all experiments demonstrated that the enzymatic isolation method implemented in the current study is appropriate. On the other hand, streptozotocin, 20 mg/kg, used as positive reference control concurrently to each assay, caused a clear increase in the mean Olive Tail Moment median value, which allows validating the current methodology.Under these experimental conditions, the in vivo rodent Comet assay demonstrated good sensitivity and good specificity: all the five renal genotoxic carcinogens were clearly detected in at least one expression period either directly or indirectly, as in the case of cisplatin: for this cross-linking agent, the significant decrease in DNA migration observed under standard electrophoresis conditions was clearly amplified when the duration of electrophoresis was increased up to 40 min. In contrast, epigenetic and nephrotoxic compounds failed to induce any signifcant increase in DNA migration. In conclusion, the in vivo rodent Comet assay performed on isolated kidney cells could be used as a tool to investigate the genotoxic potential of a test compound if neoplasic/preneoplasic changes occur after subchronic or chronic treatments, in order to determine the role of genotoxicity in tumor induction. Moreover, the epigenetic carcinogens and cytotoxic compounds displayed clearly negative responses in this study. These results allow excluding a DNA direct-acting mechanism of action and can thus suggest that a threshold exists. Therefore, the current in vivo rodent Comet assay could contribute to elucidate an epigenetic mechanism and thus, to undertake a risk assessment associated with human use, depending on the exposure level.     

Genotoxicity of municipal landfill leachate on root tips of Vicia faba

The genotoxicity of municipal landfill leachate was studied using the Vicia faba root-tip cytogenetic bioassay. Results show that landfill leachates collected in different seasons decreased the mitotic index (MI) and caused significant increases of micronucleus (MN) frequencies and anaphase aberration (AA) frequencies in a concentration-dependent manner (concentration expressed as ‘chemical oxygen demand’ measured by the method of potassium dichromate oxidation (COD_Cr)). In addition, a seasonal difference in genotoxicity induced by leachate was observed. The results confirm that leachate is a genotoxic agent in plant cells and imply that exposure to leachate in the aquatic environment may pose a potential genotoxic risk to organisms. The results also show that the V. faba cytogenetic bioassay is efficient, simple and reproducible in genotoxicity studies of leachate, and that there is a correlation between the genotoxicity and the chemical measurement (COD_Cr) of leachate.     

Genotoxic potential of marine sediments from the North Sea

The alkaline comet assay is a method for detecting DNA strand breaks and alkali labile sites in individual cells. An in vitro system was used to investigate the genotoxic potential of complex mixtures such as organic extracts of marine sediments. DNA damage was induced in leukocytes isolated from carp (Cyprius carpio) by exposure to organic sediment extracts from the North Sea or hydrogen peroxide as positive control, respectively. The minimum concentration for significant effects ranged from 1 to 40 mg sediment dry weight per milliliter assay volume. The sensitivity of the method was enhanced by using the DNA repair inhibitor, 1-beta-D-arabinofuranosylcytosine (ara C). From the results, it can be suggested that total organic carbon (TOC) as well as the different compositions of contaminants present in the sediment extracts may contribute to the genotoxic effects observed. The comet assay can be applied successfully as an in vitro bioassay for investigations on genotoxicity of marine sediment extracts.     

Genotoxicity assessment in aquatic environments under the influence of heavy metals and organic contaminants

The genotoxicity of river water and sediment including interstitial water was evaluated by microscreen phage-induction and Salmonella/microsome assays. Different processes used to fractionate the sediment sample were compared using solvents with different polarities. The results obtained for mutagenic activity using the Salmonella/microsome test were negative in the water and interstitial water samples analysed using the direct concentration method. The responses in the microscreen phage-induction assay showed the presence of genotoxic or indicative genotoxic activity for at least one water sample of each site analysed using the same concentration method. Similar results were obtained for interstitial water samples, i.e. absence of mutagenic activity in the Salmonella/microsome test and presence of genotoxic activity in the microscreen phage-induction assay. Metal contamination, as evidenced by the concentrations in stream sediments, may also help explain some of these genotoxic results. Stream sediment organic extracts showed frameshift mutagenic activity in the ether extract detected by Salmonella/microsome assay. The concentrates evaluated by microscreen phage-induction assay identified the action of organic compounds in the non-polar, medium polar and polar fractions. Thus, the microscreen phage-induction assay has proven to be a more appropriate methodology than the Salmonella/microsome test to analyse multiple pollutants in this ecosystem where both organic compounds and heavy metals are present.     

Exposure of juvenile chinook and chum salmon to chemical contaminants in the Hylebos Waterway of Commencement Bay, Tacoma, Washington

The Hylebos Waterway is an industrialized waterway ofCommencement Bay, Tacoma, Washington, that is severelycontaminated with aromatic and chlorinatedhydrocarbons in the sediment. Juvenile chinook (Oncorhynchus keta) and chum salmon (O.tshawytscha) inhabit this waterway for a few days orweeks during their outmigration from freshwaterstreams to saltwater. The purpose of thisinvestigation was to determine to what degree juvenilechum and chinook salmon captured from the HylebosWaterway might bioaccumulate organic contaminants. These levels of exposure will be compared to previousstudies where such exposures have been linked tobiological dysfunction in juvenile salmon. Theresults showed that juvenile chum and chinook salmonfrom the Hylebos Waterway take up a wide range ofchemical contaminants, compared to fish fromhatcheries or reference estuaries. These contaminantsinclude high and low molecular weight polycyclicaromatic hydrocarbons (PAHs), polychlorinatedbiphenyls (PCBs, including the toxic congeners 105 and118), hexachlorobutadiene (HCBD), hexachlorobenzene(HCB), DDTs, heptachlor, and several pesticides. Immunohistochemical examination of the gill and gut injuvenile chum salmon from the Hylebos Waterway showedthe induction of the P450 metabolizing enzyme. Moreover, concentrations of contaminants in juvenilechinook and chum salmon from the Hylebos Waterway arecomparable to levels previously shown to be associatedwith biological injury in juvenile chinook salmon,such as impaired growth, suppression of immunefunction as demonstrated by reduced B cell function,and increased mortality following pathogen exposure.     

Study of the biological impact of organic contaminants on mussels (Mytilus galloprovincialis L.) from the Venice Lagoon,Italy: responses of CYP1A-immunopositive protein and benzo(a)pyrene hydroxylase activity

A survey to evaluate the impact of organic contaminants on the mussel Mytilus galloprovincialis in the Venice Lagoon, Italy was carried out in May 1993. M. galloprovincialis were sampled from putative moderately contaminated (Alberoni, Lio Grande, Crevan), urban (Salute) and industrial (CVE) sites in the Venice Lagoon, and from a clean reference site (Plataforma) in the adjacent Adriatic Sea. Measurements comprised (i) whole-tissue body burdens of aliphatic hydrocarbons, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and other organochlorines (DDTs, hexachlorocyclohexanes and hexachlorobenzene); and (ii) digestive gland microsomal cytochrome P450 (CYP)-dependent monooxygenase system (i.e. total CYP and cytochrome P4501A (CYP1A)-immunopositive protein levels, benzo(a)pyrene hydroxylase (BPH) activity) as a specific biomarker of impact by organic contaminants. Chemical analysis identified a contaminant gradient with Plataforma as the cleanest and CVE followed by Salute as the most contaminated extremes. No elevation of total CYP content or CYP1A-immunopositive protein level was seen at any of the lagoon sites compared with Plataforma. In contrast, BPH activity and BPH turnover (i.e. BPH activity per amount total CYP) were respectively 1- and 2.5-fold higher at CVE than Plataforma (P < 0.05), and indicated to be higher (up to 1-fold) at all the other lagoon sites compared with Plataforma. Correlation was seen between BPH activity and tissue levels of total aliphatic hydrocarbons (r = 0.94-0.98), but not between the former and total PAHs or PCBs. The results are consistent with other studies in the area and indicate greatest biological impact of contaminants was at CVE followed by the other lagoon sites, with a possible genotoxic role for the elevated BPH activity in the formation of bulky DNA-adducts.     

Biomonitoring of the genotoxic potential (micronucleus and erythrocyte nuclear abnormalities assay) of the Admiralty Bay water surrounding the Brazilian Antarctic Research Station “Comandante Ferraz,” King George Island

The micronucleus (Mn) and other kinds of erythrocyte nuclear abnormality (ENA) assays were used to assess the genotoxic potential of the seawater in front of the Brazilian Antarctic Research Station “Comandante Ferraz” (King George Island). In two consecutive summers, the fish Trematomus newnesi was exposed to the seawater in front of the fuel storage tanks and the sewage discharge outlet of the Station in laboratory bioassays and in situ caging assays. After the exposure, frequencies of ENA and Mn were scored on blood smears. Significantly higher Mn and ENA frequencies in relation to those of the controls were observed in fish exposed to both types of water, in both types of experiment. Seawater in front of the fuel tanks and sewage discharge outlet at the Station was found to induce the formation of micronuclei and other ENAs, indicators of genotoxicity. The need for further investigation and the suitability of the methods for screening mutagenic pollution in polar waters are discussed.     

Comparative genotoxicity of nitrosamine drinking water disinfection byproducts in Salmonella and mammalian cells

Nitrosamine water disinfection byproducts (DBPs) are an emerging class of non-halogenated, nitrogen-containing water contaminants. Five nitrosamine DBPs were analyzed for genotoxicity (N-nitrosodimethylamine (NDMA), N-nitrosopiperidine (NPIP), N-nitrosopyrrolidine (NPYR), N-nitrosomorpholine (NMOR) and N-nitrosodiphenylamine (NDPhA). Using Salmonella typhimurium strain YG7108 the descending rank order of mutagenicity was NDMA>NPIP>NMOR>NPYR; NDPhA was not mutagenic. We developed and calibrated an exogenous S9 mix that was highly effective in activating NDMA in Chinese hamster ovary (CHO) cells using the SCGE (Comet) assay. The descending rank order for genotoxicity was NDMA>NPIP>NMOR. NDPhA was genotoxic only at one concentration and NPYR was not genotoxic. The genotoxic potencies in S. typhimurium and CHO cells were highly correlated. Based on their comparative genotoxicity attention should be focused on the generation and occurrence of NDMA, NPIP and NMOR. Current drinking water disinfection processes may need to be modified such that the generation of nitrosamine DBPs is effectively limited in order to protect the environment and the public health.     

Comparative ecotoxicity of suspended sediment in the lower Rhone River using algal fractionation,Microtox® and Daphnia magna bioassays

The toxicity associated with suspended sediments from the Rhone River (Switzerland-France) was determined with three acute bioassays. Large volume water samples were centrifuged for recovery of suspended solids in November 1989; one sample was taken as a control upstream from Lake Geneva and the 9 remainder downstream from Geneva to the Mediterranean Sea, with a single sample of the major tributary the Saône at Lyon. Heavy metals (Hg, Cd, Cr, Cu, Ni, Pb, Zn) and organic contaminants (OCs, PCBs, PAHs) bound to sediment were analysed and extracted by elutriation with filtered lake water and by organic solvent (dichloromethane). Sediment water elutriates were tested with algal fractionation bioassays (AFB) using Lake Geneva ambient phytoplankton, with Daphnia magna and Microtox® acute toxicity tests, whereas organic extracts were utilized in the latter two bioassays to evaluate the potential sediment toxicity.The bulk analyses of the sediment together with elutriate metal concentrations indicated the highest contamination of sediment downstream of Lyon. Medium contamination appeared for the stations downstream of Geneva, in the Saône River and at the Rhone River mouth. The station upstream of Lyon had low concentrations, comparable to the values in the Upper Rhone. Organic contaminants are mainly observed downstream of Lyon and their concentrations decline onwards to the sea. The bioassays Microtox® on organic extracts and AFB on the elutriates show a similar toxicity trend, but differ in that Microtox was more sensitive to organics whereas the algal test responded predominantly to metals. This difference is believed to be due to the different extraction procedures used, rather than to the tests themselves. Daphnia magna was the least sensitive and appeared to give a broader band response to the observed contaminants in the sediment. The bioassay results when integrated confirm that the biotoxicity trends relate well to the composition of the sediment, a factor which emphasizes the need for battery testing in ecotoxicological assessment.     

An Integrated Case Study for Evaluating the Impacts of an Oil Refinery Effluent on Aquatic Biota in the Delaware River: Introduction,Study Approach,and Objectives

This is the first in a series of article presenting results from a case study designed to assess the impacts of an oil refinery effluent [primarily polynuclear aromatic hydrocarbons (PAHs)] on aquatic biota in the Delaware River. During the course of the study, the oil refinery was owned by Motiva Enterprises LLC. This article provides background information on the study area, the study approach and objectives. The specific objectives of this multiyear study were to: (1) measure water column concentrations of PAHs and other contaminants (i.e., metals) in Motiva's effluent and intake canal and selected Delaware River sites; (2) assess fate and transport issues associated with the Refinery effluent; (3) characterize sediment PAHs, total organic carbon (TOC), and grain size distributions in the discharge canal, near-field, mid-field and far-field areas of the Refinery to aid in the selection of Triad sample sites (including reference areas); (4) conduct Triad studies (chemical characterizations, sediment toxicity assessments, and benthic community characterizations) at selected study sites during the spring and summer of 2001 and 2002; (5) perform fingerprinting of PAHs in Motiva's effluent to differentiate Motiva-related PAHs in sediment and biota from other sources; (6) assess bioavailability of PAHs, PCBs, and metals by using resident bivalve studies; (7) conduct long-term coring to determine potential impact of past non-complying discharges; and (8) integrate and analyze all study components to address the research goals. The results from objectives 1, 2, and 3 are briefly summarized in this series of articles whereas the other five objectives are the subject of the various papers presented in this volume.     

Cytotoxicity and genotoxicity of butyl cyclohexyl phthalate

Butyl cyclohexyl phthalate (BCP) is frequently used in personal care products, medical and household applications. The aim of this study is therefore to evaluate possible cytotoxicity and genotoxicity of BCP using in vitro and in vivo assays. The in vitro cytotoxic effect of BCP was investigated on mouse fibroblastic cell line (L929 cells) by MTT assay. The result showed that BCP inhibits cell proliferation in a concentration-dependent manner (IC₅₀ value = 0.29 µg/mL). For genotoxicity assessment, tested concentrations of BCP demonstrated mutagenic activity in the presence of S9 mix with the Salmonella strain TA100 in the Ames test. Results showed that BCP is a secondary mutagenic substance even in low concentrations. The data obtained from 28-days repeated toxicity tests on mice revealed that BCP caused abnormalities of chromosome number, in a dose-dependent manner. Additionally, DNA damage, particularly DNA strand breaks, was assessed by Comet assay. The test result shows that BCP seemed to have genotoxic potential at a high level of exposure.     

Effects of field-contaminated sediments and related water soluble components on haemocyte function and Perkinsus marinus susceptibility and expression in oysters

This paper reviews and discusses our recent findings on the effects of contaminated sediments (CSs) and related water-soluble fractions (WSFs) on haemocyte function/activity and the onset and progression of an infectious disease caused by the protozoan parasite, Perkinsus marinus (Dermo) in the eastern oyster, Crassostrea virginica. Sediments used to generate WSFs and sediments used for the whole CS exposure experiments were collected in different areas of the southern branch of the Elizabeth River, a heavily polluted sub-estuary of the Chesapeake Bay, USA. The WSFs were dominated by low molecular weight polycyclic aromatic hydrocarbons (PAHs). The CSs used for whole CS exposure experiment had elevated concentrations of high molecular weight PAHs. Polychlorinated biphenyls (PCBs) and metals were also present in the CSs. No PCBs were detected in the WSFs. In vitro exposure of haemocytes to WSFs derived from CSs reduced to haemocytes' chemotaxic, phagocytic, and chemiluminescent responses to some extent. Exposure of oysters to suspended CSs stimulated neutral red uptake, mitochondrial dehydrogenase production and ³H-leucine incorporation in haemocytes. Exposure of oysters to 0, 15, 30% WSFs increased the oysters' susceptibility to laboratory-induced infection caused by P. marinus. Exposure of oysters to 15, and 30% dilutions of WSFs for 33 days or to 1.0, 1.5, and 2.0g CSs for 30 days significantly elevated the expression/progression of latent P. marinus infection in oysters in a dose-dependent manner.     

DNA strand breaks and adducts determined in feral and caged chub (Leuciscus cephalus) exposed to rivers exhibiting variable water quality around Birmingham, UK

This study forms part of an investigation into the effects on fish of immersion in three rivers around Birmingham, UK. The rivers Blythe, Cole and Tame exhibit relatively high, intermediate and poor overall water quality, respectively, according to combined levels of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs), as well as heavy metals. Specifically, biomarkers of genotoxicity (DNA strand breaks and adducts) were measured in feral and caged chub (Leuciscus cephalus), complementing another study in which data were presented for a number of other hepatic biomarkers measured in the same animals. In both feral and caged chub, there was a general elevation of DNA strand breaks with a decrease in chemical water quality, with some time points exhibiting significantly higher levels at the most (Tame) compared with least polluted sites (Blythe), particularly in the cage-held animals. Combined-season DNA adduct data suggested a higher degree of toxic insult in the feral compared with caged chub and revealed particularly high levels of adducts in fish caught from the Cole. The pattern of adducts shown was typical of exposure to a complex mixture of PAHs which were relatively high, and similar, in both the Cole and Tame. Overall, these data are consistent with exposure of both feral and caged chub to contaminants which are able to induce specific, moderately genotoxic effects.     

Genotoxic damage in polychaetes: A study of species and cell-type sensitivities

The marine environment is becoming increasingly contaminated by environmental pollutants with the potential to damage DNA, with marine sediments acting as a sink for many of these contaminants. Understanding genotoxic responses in sediment-dwelling marine organisms, such as polychaetes, is therefore of increasing importance. This study is an exploration of species-specific and cell-specific differences in cell sensitivities to DNA-damaging agents in polychaete worms, aimed at increasing fundamental knowledge of their responses to genotoxic damage. The sensitivities of coelomocytes from three polychaetes species of high ecological relevance, i.e. the lugworm Arenicola marina, the harbour ragworm Nereis diversicolor and the king ragworm Nereis virens to genotoxic damage are compared, and differences in sensitivities of their different coelomic cell types determined by use of the comet assay. A. marina was found to be the most sensitive to genotoxic damage induced by the direct-acting mutagen methyl methanesulfonate (MMS), and showed dose-dependent responses to MMS and the polycyclic aromatic hydrocarbon benzo(a)pyrene. Significant differences in sensitivity were also measured for the different types of coelomocyte. Eleocytes were more sensitive to induction of DNA damage than amoebocytes in both N. virens and N. diversicolor. Spermatozoa from A. marina showed significant DNA damage following in vitro exposure to MMS, but were less sensitive to DNA damage than coelomocytes. This investigation has clearly demonstrated that different cell types within the same species and different species within the polychaetes show significantly different responses to genotoxic insult. These findings are discussed in terms of the relationship between cell function and sensitivity and their implications for the use of polychaetes in environmental genotoxicity studies.     

Genotoxicity of hydroquinone in A549 cells

Hydroquinone (HQ) is found in natural and anthropogenic sources including food, cosmetics, cigarette smoke, and industrial products. In addition to ingestion and dermal absorption, human exposure to HQ may also occur by inhaling cigarette smoke or polluted air. The adverse effects of HQ on respiratory systems have been studied, but genotoxicity HQ on human lung cells is unclear. The aim of this study was to investigate the cytotoxicity and genotoxicity of HQ in human lung alveolar epithelial cells (A549). We found that HQ induced a dose response in cell growth inhibition and DNA damage which was associated with an increase in oxidative stress. Cytotoxicity results demonstrated that HQ was most toxic after 24 h (LC₅₀?=?33 μM) and less toxic after 1 h exposure (LC₅₀?=?59 μM). Genotoxicity of HQ was measured using the Comet assay, H2AX phosphorylation, and chromosome aberration formation. Results from the comet assay revealed that DNA damage was highest during the earlier hours of exposure (1 and 6 h) and thereafter was reduced. A similar pattern was observed for H2AX phosphorylation suggesting that damage DNA may be repaired in later exposure hours. An increase in chromosomal aberration corresponded with maximal DNA damage which further confirmed the genotoxic effects of HQ. To investigate whether oxidative stress was involved in the cytotoxic and genotoxic effects of HQ, cellular glutathione and 8-Oxo-deoguanisone (8-Oxo-dG) formation were measured. A decrease in the reduced glutathione (GSH) and an increase oxidized glutathione (GSSG) was observed during the early hours of exposure which corresponded with elevated 8-Oxo-dG adducts. Together these results demonstrate that HQ exerts its cytotoxic and genotoxic effects in A549 lung cells, probably through DNA damage via oxidative stress.     

Transfer of microcontaminants from sediment to chironomids, and the risk for the Pond bat Myotis dasycneme (Chiroptera) preying on them

Transfer ratios of metals, PCBs, pesticides and PAHs from the sediment to chironomid larvae and adults collected in a highly contaminated area, the Biesbosch, were studied. Metal concentrations in larvae were 0.28 (Cd), 0.02 (Cr), 0.52 (Cu), 0.06 (Hg), 0.03 (Pb), 0.32 (Zn) times those found in standard sediment, on a dry weight basis. Hg and Zn were well transported to the adult stage. Dry weight ratios of contaminant residues in adults and in larvae were 0.38 (Cd), 0.23 (Cr), 0.62 (Cu), 1.03 (Hg), 0.08 (Pb), 0.94 (Zn). For PCBs and pesticides, the concentration ratios of chironomid larvae fat to sediment (dry organic matter) varied around 3.3, which is consistent with laboratory studies. Organochlorine residues in adult fat were comparable to those in larvae lipids. However, the concentrations of non-ortho PCBs were 1.7 times higher in adults. For polycyclic aromatic hydrocarbons (PAH), larval fat:sediment (organic matter) concentration ratios ranged from 0.004 to 0.1. Adult: larvae ratios for PAHs varied between 0.2 and 0.6. For naphthalene a much higher transport ratio of 2 was found. Chironomid adults are the most important potential food source of the Pond bat, which lives in low densities in the Biesbosch. The contaminant concentrations measured in the chironomids do not exceed diet levels that are thought to be safe for mammals. However, Pond bats collected in less contaminated areas contained PCB-concentrations of 9, 33 and 76 mg kg^–1 lipid weight, which are above concentrations that cause reproduction effects on Mink.     

Distribution,Sources Identification,and Ecological Risk of PAHs and PCBs in Coastal Surface Sediments from the Northern Persian Gulf

Hormozgan Province plays a vital role in fishery, petroleum, and industrial activities in southern Iran. However, no comprehensive studies on organic pollution have been performed. PCBs and PAHs were analyzed in surface sediments from areas receiving industrial (nine sites), river (one site), and urban (two sites) effluents. The sediment samples were collected in March and September 2010 (in dry and wet seasons) at the highest tidal time. The overall pollution level of PCBs ranged from 2.5 ± 0.8 to 462.0 ± 206.7 ng/g dry weight. CB153 congener dominated in most of the sediment samples. Congener profiles of PCBs showed close similarity with formulations of commercial products such as Aroclor 1260 and 1254 g. A wide range of 55.3 to 1231.6 ng/g dry weight was detected for ∑PAHs. Results of PCA and PCA-MLR tests confirmed both petrogenic and pyrogenic origins for PAH pollution. The higher means of ∑PAHs and ∑PCBs in industrial and urban wastewaters were found near the shore, evidencing the role of these wastewaters in the PAH and PCB contamination in Hormozgan sediment. The concentrations of PAHs and PCBs in detected hotspots exceed the U.S. NOAA sediment quality guidelines.     

Genotoxicity is modulated by ascorbic acid: Studies using the wing spot test in Drosophila

The ability of ascorbic acid (Vitamin C) to modulate the genotoxic action of several mutagens was investigated in the wing spot test of Drosophila melanogaster. In this assay, 3-day-old transheterozygous larvae for the multiple wing hairs (mwh, 3-0.3) and flare (flr, 3-38.8) genes were treated with three reference mutagenic compounds, namely cobalt chloride (CoCl₂), 4-nitroquinoline 1-oxide (4-NQO) and potassium dichromate (K₂Cr₂O₇). The results obtained show that the three reference mutagens tested were clearly genotoxic in the Drosophila wing somatic mutation and recombination test (SMART). None of the three concentrations tested of ascorbic acid (25, 75 and 250 mM) induced significant increases in the frequency of the mutant clones recorded. When co-treatment experiments with ascorbic acid were carried out, different results were found. Thus, ascorbic acid was effective in reducing the genotoxicity of K₂Cr₂O₇ virtually to the control level; on the contrary, it did not show any antigenotoxic effect on the genotoxicity of 4-NQO. Finally, co-treatments with CoCl₂ and ascorbic acid show a significant increase in the frequency of mutant clones over the values obtained with CoCl₂ alone.